6种鱼类病原菌的免疫反应分析及其检测免疫芯片的构建  

Construction of Immunochip for Detection of Six Fish Bacterial Pathogens

作　　者：李永芹[1] 绳秀珍[1] 战文斌[1] 徐晓丽[1] 

机构地区：[1]中国海洋大学海水养殖教育部重点实验室,青岛266003

出　　处：《中国海洋大学学报（自然科学版）》2010年第8期48-54,共7页Periodical of Ocean University of China

基　　金：国家高技术研究发展计划项目(2006AA100306);国家自然科学基金项目(30771648)资助

摘　　要：采用ELISA、Western-blot及Dot-blot方法,分析海豚链球菌(Streptococcus iniae)、鳗弧菌(Vibrioanguillarum)、杀鲑气单胞菌(Aeromonas salmonicida)、迟缓爱德华氏菌(Edwardsiella tarda)、荧光假单胞菌(Psedo-monas fluorescens)及海分支杆菌(Mycobacterium marinum)6种养殖鱼类病原菌与其兔抗血清之间的免疫反应。结果显示,ELISA、Western-blot和Dot-blot 3种方法的分析结果具有一致性,各菌株抗血清与相应的抗原菌株间的反应最为强烈,各菌株与其他细菌兔抗血清间有程度不等的交叉反应。基于以上结果,采用6种病原菌的兔抗血清作为捕获抗体构建了其免疫芯片,确定了检测结果的判读方法,并对其进行验证性应用,结果显示,该芯片可以用于病原菌的准确检测,对分别注射感染不同病原菌的牙鲆(Paralichthys olivaceus)进行取样检测的结果也验证了这一结论。ELISA,Western-blot and Dot-blot were used to analyze the cross reactions between 6 strains of pathogenic bacteria（including Streptococcus iniae,Vibrio anguillarum,Aeromonas salmonicida,Edwardsiella tarda,Psedomonas fluorescens and Mycobacterium marinum） and their antisera.The results obtained by three methods all showed that the reactions between the pathogenic bacteria and their own antiserum were the strongest,and there were different levels of cross reactions between the antisera of S.iniae,A.salmonicida,P.fluorescens,M.marinum and others bacteria respectively.Basing on above results,an immunochip for bacterial detection was developed,using the antisera against 6 strains of pathogenic bacteria as capture antibody respectively;and the determination method of the bacteria-detecting results was confirmed.Finally the integrated immunochip was applied in the detection of 6 strains of bacteria,and the results revealed that the pathogenic bacteria could be detected accurately in spite of the cross reactions in existence,which was further confirmed by the detection of infected tissues from Japanese flounder（Paralichthys olivaceus） injected with E.tarda or the mixture of V.anguillarum,P.fluorescens and M.marinum.

关 键 词：ELISA WESTERN-BLOT DOT-BLOT 免疫芯片 交叉反应 

分 类 号：S94[农业科学—水产养殖]